Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Stat3 (Ser727) Antibody #9134

Acute-phase response factor   APRF   pstat3   sc-8001   signal transducer and activator of transcription  

No. Size Price
9134L 300 µl ( 30 western blots ) ¥8,792.00 现货查询 购买询价
9134S 100 µl ( 10 western blots ) ¥3,900.00 现货查询 购买询价
9134T 20 µl ( 2 western blots ) ¥1,500.00 现货查询 购买询价
9134 carrier free & custom formulation / quantityemail request
Applications Dilution Species-Reactivity Sensitivity MW (kDa) Isotype
W 1:1000 Human,Mouse,Rat, Endogenous 86 Rabbit
IP 1:50
ChIP 1:50

Species cross-reactivity is determined by western blot.

Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP,

Homology

Species predicted to react based on 100% sequence homology: Hamster, Bovine, Dog, Pig, Horse,

Specificity / Sensitivity

Phospho-Stat3 (Ser727) Antibody detects endogenous levels of Stat3 only when phosphorylated at Ser727. The antibody does not significantly cross-react with the corresponding phospho-serines of other Stat proteins.

Phospho-Stat3 (Ser727) Antibody只能检测内源的在Ser727位点磷酸的Stat3 蛋白。此抗体不与在对应丝氨酸残基磷酸化的其它Stat 蛋白交叉反应。

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser727 of mouse Stat3. Antibodies are purified by protein A and peptide affinity chromatography.

Phospho-Stat3 (Ser727)多克隆抗体是通过合成鼠源对应的Stat3 Ser727位点周围的磷肽段来免疫动物而获得。抗体是通过protein A和多肽亲和层析法纯化。

Western Blotting

Western Blotting

Western blot analysis of extracts from A431 cells, untreated or EGF-treated (100 ng/ml) for the indicated times, using Phospho-Stat3 (Ser727) Antibody (upper) or Stat3 Antibody #9132 (lower).Western免疫印迹。用Phospho-Stat3 (Ser727) Antibody (上图) 或 Stat3 Antibody #9132 (下图)研究未经处理或经EGF处理(100 ng/ml) 一定时间的A431细胞的细胞抽提液。

IF-IC

IF-IC

Confocal immunofluorescent analysis of HeLa cells, untreated (left), IL-6 treated (center) or IL-6 and phosphatase treated (right), labeled with Phospho-Stat3 (Ser727) Antibody (green) and Pan-Keratin (C11) Mouse mAb #4545 (red).共聚焦免疫荧光分析未经处理的HeLa(左图)或者经IL-6 处理的 HeLa (中图)或经过IL-6和 磷酸酶处理的HeLa 细胞 (右图) ,所用抗体为Phospho-Stat3 (Ser727) Antibody (绿色)和 Pan-Keratin (C11) Mouse mAb #4545 (红色)。

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Hep G2 cells starved overnight and treated with IL-6 (100 ng/ml) for 30 minutes, and either 20 μl of Phospho-Stat3 (Ser727) Antibody or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human IRF-1 promoter primers, SimpleChIP® Human c-Fos Promoter Primers #4663, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.染色质免疫共沉淀。Hep G2 细胞培养至4 x 106, 并经过夜饥饿和IL-6(100 ng/ml,30 min) 处理,然后用SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003进行免疫沉淀实验,本实验中用20 μl Phospho-Stat3 (Ser727) Antibody或2 μl Normal Rabbit IgG #2729 抗体。用人源 IRF-1 promoter primers, SimpleChIP® Human c-Fos Promoter Primers #4663, and SimpleChIP® Human α Satellite Repeat Primers #4486对富集的DNA做real-time PCR。每个样本中沉淀的DNA量定义为相对信号与输入的总染色质相比的数值。

Western Blotting

Western Blotting

Western blot analysis of extracts from A172 cells, untreated (-) or UV-treated (100 mJ, 30 min; +) with or without λ phosphatase (+), using Phospho-Stat3 (Ser727) Antibody (upper) and Stat3 (79D7) Rabbit mAb #4904 (lower).

Background

The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).

Stat3 转录因子是细胞因子和生长因子受体信号通路上重要的信号分子(1),并且被鼠科胚胎的发育所需要(2)。Stat3 在很多的人类肿瘤中是组成性激活的(3,4)并有潜在的支配原发性癌的可能(5) 而且也具有抗细胞凋亡的活性(3)。Stat3通过Tyr705位点的磷酸化而激活,从而导致它的二聚化,入核和DNA结合(6,7)。转录活性似乎受到 MAPK 或 mTOR 信号通路介导的Stat3 Ser727位点的磷酸化(8,9)。Stat3不同剪切形式的表达反应了其生物学功能,Stat3α (86 kDa) 和 Stat3β (79 kDa) 形式的表达比例依赖于细胞类型,配体方向或者细胞发育的成熟阶段(10)。值得注意的是Stat3β蛋白在C-末端转录活性区域缺少丝氨酸磷酸化位点(8)。

  1. Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
  2. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
  3. Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
  4. Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
  5. Bromberg, J.F. et al. (1999) Cell 98, 295-303.
  6. Darnell, J.E. et al. (1994) Science 264, 1415-21.
  7. Ihle, J.N. (1995) Nature 377, 591-4.
  8. Wen, Z. et al. (1995) Cell 82, 241-50.
  9. Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
  10. Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.

Application References

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For Research Use Only. Not For Use In Diagnostic Procedures.

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Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.

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